表皮调节素、上皮有丝分裂原和干细胞因子对山羊精原干细胞增殖的影响

Effect of Epiregulin,Epigen and SCF on Proliferationof Goat Spermatogonial Stem Cells

为了揭示表皮调节素(Epiregulin/EREG)、上皮有丝分裂原(Epigen/EPG)和干细胞因子(SCF)对山羊精原干细胞(SSCs)增殖的影响,改进SSCs体外培养体系。在含有胶质细胞源性神经营养因子(GDNF)、碱性成纤维细胞生长因子(bFGF)和白细胞抑制因子(LIF)的SSCs培养基基础上,分别添加20 ng/mL干细胞因子、20 ng/mL表皮调节素、20 ng/mL和100 ng/mL上皮有丝分裂原。对富集后山羊SSCs进行12 d的培养,测定培养过程中山羊SSCs生长曲线和细胞活性。结果表明:添加100 ng/mL上皮有丝分裂原和20 ng/mL的表皮调节素相比其他组能够显著促进山羊SSCs的体外增殖和细胞活力(P<0.05);其中培养12 d后添加20 ng/mL表皮调节素的细胞活力显著高于其他组,但细胞数量与100 ng/mL上皮有丝分裂原组无显著差异。20 ng/mL SCF相比对照组能够显著提高体外增殖和细胞活力(P<0.05),但低于100 ng/mL上皮有丝分裂原和20 ng/mL表皮调节素组(P>0.05)。因此,添加100 ng/mL上皮有丝分裂原、20 ng/mL干细胞因子和20 ng/mL表皮调节素能够提高山羊SSCs的增殖,其中添加20 ng/mL表皮调节素效果最佳。

In order to study the effects of Epiregulin(EREG),Epigen(EPG)and Stem Cell Factor(SCF)on the proliferation of goat spermatogonial stem cells(SSCs)and to improve SSCs culture system,this experiment added 20 ng/mL SCF,20 ng/mL EREG,20 ng/mL,and 100 ng/mL EPG in the spermatogonial stem cell culture medium containing FBS,GDNF,bFGF,and LIF.Then the enriched goat SSCs were cultivated for 12 days.The growth curve and cell activity of goat SSCs during culture were measured.The results showed that the groups treated with 100 ng/mL and 20 ng/mL EPG can significantly(P<0.05)increase the in vitro proliferation and cell viability of goat SSCs.After 12 days culture,the cell vitality of 20 ng/mL EREG was significantly higher than the control group(P<0.05),and there was no difference in the cell number with the 100 ng/mL EPG group significantly(P>0.05).Compared with the control group,the group with 20 ng/mL SCF can significantly increase the proliferation and cell viability(P<0.05),but the effects were lower than the 100 ng/mL EPG and 20 ng/mL EREG groups(P>0.05).In conclusion,adding 100 ng/mL EPG,20 ng/mL SCF,and 20 ng/mL EREG can increase goat SSCs proliferation,and the 20 ng/mL EREG has the best promotion effect.